JP2005098704A - Method for fractionating particulate of different specific gravity - Google Patents

Method for fractionating particulate of different specific gravity Download PDF

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JP2005098704A
JP2005098704A JP2001070651A JP2001070651A JP2005098704A JP 2005098704 A JP2005098704 A JP 2005098704A JP 2001070651 A JP2001070651 A JP 2001070651A JP 2001070651 A JP2001070651 A JP 2001070651A JP 2005098704 A JP2005098704 A JP 2005098704A
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specific gravity
solid inclusions
particulates
method
centrifuge tube
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Hajime Ogata
肇 緒方
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Hajime Ogata
肇 緒方
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483Physical analysis of biological material
    • G01N33/487Physical analysis of biological material of liquid biological material

Abstract

PROBLEM TO BE SOLVED: To provide a method for precisely fractionating particulates in a liquid containing two types of particulates of different specific gravities or more into two groups according to their specific gravity differences by a simple method in the case as of fractionating erythrocytes from blood.
SOLUTION: Groups of particulates containing the two types of particulates having different specific gravities or more are centrifuged with a solid inclusion, having a specific gravity in an intermediate zone between the specific gravities of the groups of particulates and vertically movable with the external edge of its contact surface with the groups of particulates inscribed with a centrifuge tube, to move the particulates having a high specific gravity downward below the solid inclusion or the particulates having a low specific gravity upward over the solid inclusion through the contact surface between the inner wall of the centrifuge tube and the solid inclusion and simultaneously move the solid inclusion to an intermediate position between the groups of particulates having high and low specific gravities. The particulates having different specific gravities are thereby fractionated.
COPYRIGHT: (C)2005,JPO&NCIPI

Description

【0001】 [0001]
【発明の属する技術分野】 BACKGROUND OF THE INVENTION
本発明は、二種類の微粒子を比重の違いにより簡便かつ高い精度で分別する方法およびその装置に関する。 The present invention relates to a method and apparatus for separating simply and at high accuracy by the specific gravity difference between the two types of fine particles. 特に体液から赤血球を分画除去するのに適した方法並びに装置に関する。 To a method and apparatus suitable for fractionating remove erythrocytes from body fluids.
【0002】 [0002]
【従来の技術】 BACKGROUND OF THE INVENTION
血液などからの赤血球の除去法および有核細胞の分離法には、従来から(1)バフィー・コート採取法、(2)溶血による赤血球除去法、(3)赤血球凝集沈降分離法、(4)比重溶液の密度勾配を利用した遠心分離法などの手法が知られている。 The removal method and the nucleated cell sorting of red blood cells, such as from blood, conventionally (1) Buffy coat collection method, (2) red blood cells removal method by hemolysis, (3) Hemagglutination sedimentation method, (4) techniques such as centrifugation method using a density gradient of specific gravity solution is known.
【0003】 [0003]
【発明が解決しようとする課題】 [Problems that the Invention is to Solve
前記公知の手法には、それぞれ次のような問題点があった。 Wherein the known method, there is respectively the following problems.
(1)バフィー・コート採取法この方法は、採取しようとする有核細胞への大量の赤血球の混入が不可避で、また有核細胞の回収率も手技を施す人の熟練度に大きく左右される。 (1) buffy coat collection method this method, contamination of large amounts of red blood cells of the nucleated cells to be collected is inevitable, also largely depends on the people of proficiency to perform the recovery rate also procedure of nucleated cells .
(2)溶血による赤血球除去法この方法は、溶血に用いる試薬の細胞毒性が不可避であり、有核細胞の回収率および純度は、試薬の濃度や混合比率および反応時間といった条件に左右される。 (2) The method erythrocyte removal method by hemolysis, cytotoxicity of the reagent used in the hemolysis is unavoidable, recovery and purity of the nucleated cells will depend on the conditions such as the concentration and the mixing ratio and the reaction time of the reagents. また、大容量の検体には不向きである。 In addition, it is not suitable for specimen of the large capacity.
(3)赤血球凝集沈降分離法この方法によって満足しうる回収率を得るためには、試料を等張液で希釈するか、または擬集沈降手技を数回反復する必要がある。 (3) to obtain the erythrocyte aggregation sedimentation separation method recoveries that can be satisfied by this method, or diluted by isotonic solution sample, or it is necessary to repeat several times the agglomerative precipitation procedure. 従って、分画後に浮遊細胞を再度遠沈洗浄してデキストランやヘタスターチなどの凝集薬を除去し、多量の溶液中に拡散した状態で存在する有核細胞を回収する必要があり、作業工程が多く煩雑である。 Thus, the floating cells again far precipitation cleaned after fractionation to remove any coagulum agents such as dextran and hetastarch, it is necessary to recover the nucleated cells present in a state of being diffused in a large amount of solution, many working steps it is complicated.
(4)比重溶液の密度勾配を利用した遠心分離法この方法は、溶液の重層や目的とする比重帯の分離手技等、慎重な操作と熟練が要求される。 (4) centrifugation method This method of density gradient using specific gravity solution, separation procedures specific gravity range of the layer or object such as a solution, careful manipulation and skill is required. また、分画後に浮遊細胞を再度遠沈洗浄して、たとえばファイコール(Ficall)、パーコール(Percoll)等の比重溶液を除去する必要があり、作業工程が多く煩雑である。 Also, the floating cells were far precipitation washed again after fractionation, e.g. Ficoll (FiCall), it is necessary to remove the specific gravity solution such as Percoll (Percoll), it is troublesome that many working steps.
また、血清分離用には既にゾル状の介在物を用いて、血球等の分離を行う方法も知られてはいるが、細胞分離用として用いるためには、介在物の厳密な比重調整が必要で、かつ検体の比重分布そのものの個体差に影響されるため、安定した結果が得られない。 Moreover, the sera separated already using sol inclusions, although the method is also known to separate such blood cells, for use as a cell separation requires strict weighting of inclusions in, and since it is affected by the individual difference of the specific gravity distribution itself of the sample, not stable results.
【0004】 [0004]
【課題を解決するための手段】 In order to solve the problems]
本発明者は、比重の異なる2種以上の微粒子と特定の比重と形状を有する固体介在物を一緒に遠心することにより、高比重の微粒子を固体介在物の下方に、または低比重の微粒子を固体介在物の上方に移動させ、比重の異なる微粒子を互いに分離する方法を見つけた。 The present inventor has found that by centrifugation of the solid inclusions having two or more fine particles with a specific gravity and shape of different specific gravities together, the high specific gravity of the particles under the solid inclusions, or the low specific gravity of the fine particles is moved over the solid inclusions, they found a method for separating fine particles with different specific gravities from each other.
すなわち、本発明は、 That is, the present invention is,
(1)比重の異なる二種以上の微粒子を含む微粒子群を、その微粒子群の比重の中間帯に位置する比重を持ち、微粒子群との接触面の外縁が遠心管に内接して上下に移動可能な固体介在物と共に遠心して、遠心管内壁と固体介在物との接触面を通して高比重微粒子を固体介在物の下方に、または低比重微粒子を固体介在物の上方に移動させると同時に固体介在物を比重の高低二粒子群の中間に移動、介在させることを特徴とする比重の異なる微粒子の分別法。 (1) Move the group of fine particles comprising different two or more kinds of fine particles specific gravity, it has a specific gravity which is located in the middle zone of the specific gravity of the microparticle group, the upper and lower outer edges of the contact surface between the particulates group is inscribed in a centrifuge tube by centrifugation with possible solid inclusions, high specific gravity particles to below the solid inclusions, or low-density particulate solid inclusions simultaneously move above the solid inclusions through the contact surface between the centrifuge tube wall and the solid inclusions fractionation of fine particles having different specific gravities, characterized in that the movement in the middle of the high and low two particle groups having a specific gravity, is interposed.
(2)固体介在物の形状が、球状、楕円体状または微粒子群との接触面の少なくとも一部が遠心管内壁に対して鋭角で接触している柱状であることを特徴とする(1)記載の方法。 (2) the shape of the solid inclusions spherical, wherein at least a portion of the contact surface with the ellipsoidal or particulate groups are columnar in contact at an acute angle to centrifuge tube walls (1) the method described.
(3)微粒子の粒径が0.01〜200μmの範囲内にあることを特徴とする(1)記載の方法。 (3) the particle size of the fine particles is characterized in that in the range of 0.01~200Myuemu (1) The method according.
(4)微粒子が液体中に懸濁しているかまたはコロイド状として存在していることを特徴とする(1)記載の方法。 (4) fine particles are characterized by being present as either or colloidal suspended in a liquid (1) The method according.
(5)微粒子を含む液体が、体液であることを特徴とする(1)記載の方法。 (5) a liquid containing fine particles, characterized in that a body fluid (1) The method according.
(6)固体介在物が、寒天、ゼラチン、合成樹脂、セラミクス、タンパク質、多糖類から選ばれた一種であることを特徴とする(1)記載の方法。 (6) Solid inclusions, agar, gelatin, synthetic resin, ceramic, a protein, methods according to (1) wherein it is a one selected from a polysaccharide.
(7)遠心管内に、外縁が遠心管に内接し且つ上下移動が可能な固体介在物を収容したことを特徴とする(1)記載の方法を実施するための装置、 (7) to the centrifuge tube, outer edge and wherein the housing the inscribed and vertical movement can be solid inclusions into a centrifuge tube (1) device for carrying out the method according,
である。 It is.
【0005】 [0005]
【発明の実施の形態】 DETAILED DESCRIPTION OF THE INVENTION
本発明において、分別の対象となる比重の異なる二種以上の微粒子とは、微粒子径が0.01〜200μm程度のもので、微粒子の種類、形状を問わず、比重の異なる二種以上の微粒子を含む微粒子群、特に微粒子が液体媒体中混在している状態の微粒子群である。 In the present invention, different two or more kinds of fine particles in specific gravity to be separated, but fine diameter of about 0.01~200Myuemu, the type of particulate, regardless of the shape, different two or more kinds of fine particles specific gravity particle group including, in particular microparticles group state particles are mixed in a liquid medium. その代表的なものは、体液、特に赤血球、白血球などの種々の比重の異なる細胞を含む血液である。 Typical ones, body fluid is blood, especially including red blood cells, a variety of specific gravities of different cells, such as leukocytes. その他、比重の異なるコロイド微粒子を含むコロイド液、ミセルを含む液、、エマルション、サスペンジョンなども含まれる。 Other, colloidal solution containing a different colloid particles of specific gravity, liquid ,, emulsion containing micelles, suspension, etc. are also included. 本発明においては、特に体液中の細胞集団から赤血球を分離除去して有核細胞を採取するのに適している。 In the present invention, it is particularly suitable for collecting red blood cells from the cell population separated and removed nucleated cells in the body fluid.
本発明に用いられる遠心管1は、材質、大きさ等は、特に限定されるものではなく、通常の遠心分離器に用いられるものであればよいが、半球状、半楕円体状、コーン状などの形状を有する底部6と、それに続く円筒状の胴部7からなるものがよい。 Tube 1 used in the present invention, the material, size and the like, not limited in particular, but as long as it is commonly employed in the centrifugal separator, hemispherical, semi-ellipsoidal, conical a bottom portion 6 having a shape such as, good consist cylindrical barrel portion 7 that follows. 遠心管の材質は、ガラス、合成樹脂、ゴム、金属等が用いられる。 The material of the centrifuge tube, glass, synthetic resin, rubber, metal or the like is used.
【0006】 [0006]
本発明に用いられる固体介在物2は、比重の異なる二種以上の微粒子が、液体に分散されている場合は、その液体に不溶のもので、遠心力が加わったとき微粒子が遠心管器壁と内接している固体介在物との間を移動しうる僅かの隙間を有しているものか、または、そのような間隙を生む程度の柔軟性を有しているもの、すなわち、弾性体がよい。 Solid inclusions 2 used in the present invention, two or more particles having different specific gravity, if it is dispersed in the liquid, those insoluble in the liquid, particles centrifuge tube device wall when the centrifugal force is applied or not have a small gap that can move between the solid inclusions are inscribed, or, as it has flexibility enough to produce such a gap, i.e., the elastic member good.
固体介在物2の材質は、たとえば、寒天、ゼラチン、シリコン樹脂、ポリスチレン樹脂、ABS樹脂,ポリウレタン樹脂等の合成樹脂、ガラスなどのセラミクス、タンパク質、多糖類等が挙げられる。 The material of the solid inclusions 2, for example, agar, gelatin, silicone resin, polystyrene resin, ABS resin, synthetic resin such as polyurethane resin, ceramics such as glass, proteins, and polysaccharides.
固体介在物2の比重は、その中に比重の異なる微粉末たとえば、シリカ、クレー、セラミクスパウダー,金属粉末などを必要量混合することにより調節することができる。 Specific gravity of the solid inclusions 2, fine powders different in specific gravity therein for example, can be adjusted silica, clay, ceramic powder, by mixing a necessary amount and metal powder.
【0007】 [0007]
本発明の特徴の一つは、固体介在物を微粒子群中に介在させ、固体介在物の微粒子群との分界面の形状を遠心力方向に直交する平面以外の形状、例えば、球状、楕円体状または微粒子群との接触面の少なくとも一部が遠心管内壁に対して鋭角で接触している柱状に設定したことである。 One feature of the present invention, the solid inclusions are interposed in the microparticles group, a shape other than plane orthogonal minute surface shape of the particulate group the centrifugal force direction of the solid inclusions, for example, spherical, ellipsoidal Jo, or at least part of the contact surface between the particulate group is that set in columnar in contact at an acute angle to centrifuge tube wall.
図4〜6に示したように、遠心力により、微粒子群を含む液10の微粒子が整然とした比重勾配を形成する。 As shown in FIGS. 4-6, by centrifugal force, the fine particles of the liquid 10 containing the particles group to form an orderly density gradient. ついで固体介在物2よりも高比重の微粒子3が遠心管内壁と固体介在物との接触面9の間を擦り抜けて順々に固体介在物の下方に移動し、その結果固体介在物2は上方に移動する。 Then moved downward solid inclusions one after the other slip through between the contact surface 9 with high specific gravity particles 3 centrifuge tube walls and solid inclusions than solid inclusions 2, resulting solid inclusions 2 It moves upward. 最終的な定常状態においては、固体介在物の面8と遠心管内壁との漸近により構成される狭い空間5に微粒子群は少量の比重勾配を残したまま拮抗する。 In the final steady-state, particulate group in a narrow space 5 formed by asymptotic between the surface 8 and the centrifuge tube wall of the solid inclusions antagonize leaving a small amount of density gradient. 従って、微粒子群自体の比重にロット差がある場合でも、目的とする低比重微粒子4を他の微粒子の混在が少ない状態で安定して分別することができる。 Therefore, even if there is lot difference specific gravity of the group of fine particles per se, it is possible to stably fractionate the low specific gravity particles 4 of interest in a state mixed with less other particulate.
固体介在物2の望ましい形状の一つは、球(図3)または楕円体である。 One preferred shape of the solid inclusions 2 is a sphere (FIG. 3) or an ellipsoid. 他の望ましい形状は、固体介在物が微粒子群に接している面と遠心力方向、すなわち遠心管内壁とのなす角度αが鋭角である柱状体(図1および2)である。 Other desirable shapes is a surface and the centrifugal force direction which the solid inclusions are in contact with the group of fine particles, i.e. columnar bodies angle α is an acute angle between the centrifuge tube wall (Fig. 1 and 2). この場合の角度は、20〜70度が好ましく、30〜60度であることがより好ましい。 Angle in this case is preferably 20 to 70 degrees, and more preferably 30 to 60 degrees.
固体介在物が微粒子と接する面8は、平面であっても、曲面であってもよい。 Surface 8 solid inclusions are in contact with the fine particle may be planar or may be curved. 固体介在物の比重は、微粒子分別の目的に応じて適宜選択することができるが、分別しようとする二つの微粒子の比重の間にくるように適当な比重を有する物質を混入することで、その比重を任意に調節することができる。 Specific gravity of the solid inclusions, it may be appropriately selected depending on the purpose of the microparticle fractionation, to incorporate a material having a suitable density to come between the specific gravity of the two particles to be separated, their it is possible to adjust the specific gravity arbitrarily.
【0008】 [0008]
遠心管底部内壁には、遠心した際、固体介在物が遠心管の上下に移動しやすくするために、パラフィン等の離型剤を塗布しておくこともできる。 The centrifuge tube bottom inside wall, upon centrifugation, for solid inclusions to easily move up and down the tubes, it is also possible to keep applying a release agent such as paraffin.
次に本発明の代表的な実施方法について、図面を参照しながら説明する。 Next exemplary method of the present invention will be described with reference to the drawings.
まず、図4に示される通り、半径rの半球状の底部6とそれに続く円筒状胴部7を有する遠心管1に、上面に傾斜面8を有する固体介在物2を、遠心管1に内接するように配置する。 First, the inner as shown in FIG. 4, the tube 1 having a cylindrical barrel 7 followed by a hemispherical bottom 6 of the radius r, the solid inclusions 2 having an inclined surface 8 on the upper surface, the centrifuge tube 1 placing in contact. この遠心管1に赤血球3(高比重微粒子)と有核細胞4(低比重微粒子)の比重の異なる二種の微粒子を含む血液10を入れ、遠心管を遠心機にかける。 The centrifuge tube 1 placed blood 10 that contains the two kinds of particles having different specific gravities of erythrocytes 3 (high density particles) and nucleated cells 4 (low density particles), applying a centrifugal tube centrifuge. 遠心を始めると、高比重である赤血球3が固体介在物傾斜面と遠心管内壁が漸近してくる部分5に集まり、さらに遠心力が加わるとその赤血球3は、遠心管内壁と固体介在物との間5を擦り抜けて固体介在物の下方に移動し、一方固形介在物2は、赤血球との比重差により生じる負の遠心力により押し上げられて上方に移動する。 Starting centrifugation, red blood cells 3 is a high specific gravity gather in the portion 5 of the solid inclusions inclined surface and the centrifuge tube wall comes asymptotically, further centrifugal force is applied the red blood cells 3, the centrifuge tube wall and solid inclusions exits rubbed 5 between moves downwards of the solid inclusions, whereas solid inclusions 2 moves upwardly pushed up by the negative of the centrifugal force resulting from the difference in specific gravity between the red blood cells. 遠心終了時点では、血球は低比重から高比重へと整然と積層する。 The centrifugal end, blood cells stacked neatly to high density low density. そして、固体介在物の下方には大部分の赤血球3が移動し、有核細胞4と赤血球3とが接する界面9に固体介在物2が介在する。 Then, on the lower side of the solid inclusions move most of the red blood cells 3, the solid inclusions 2 is interposed at the interface 9 in contact and the nucleated cells 4 and erythrocytes 3.
比重の異なる二種以上の微粒子の混合物10は、固体介在物の下方に配置してもよい。 Mixtures of different two or more particulate specific gravity 10 may be disposed below the solid inclusions. この場合は、まず、遠心管に、比重の異なる二種以上の微粒子の混合物10を入れ、その上に成形された固体介在物2を挿入する。 In this case, first, a centrifuge tube, the mixture was placed 10 of two or more particles having different specific gravities, inserting the solid inclusions 2 molded thereon. この場合の固体介在物の形状は,球形または楕円形のものが、挿入が容易であることにおいて好ましい。 The shape of the solid inclusions in this case is preferred in that those spherical or oval, it is easy to insert. この比重の異なる二種以上の微粒子の混合物と固体介在物を収容した遠心管を遠心すると低比重微粒子4が遠心管内壁と固体介在物との接触面の間9を擦り抜けて固体介在物の上方に移動し、高比重粒子3が固体介在物の下方に留まる。 Mixtures and solid inclusions centrifuged accommodating the centrifugal tube a low specific gravity particles 4 is missing rubbing between 9 of the contact surface between the centrifuge tube wall and the solid inclusions solid inclusions of two or more particles having different this specific gravities moves upward, the high specific gravity particles 3 remain below the solid inclusions. その際、固体介在物2は、低比重粒子4が上方に移動した分だけ下方に移動する。 At that time, the solid inclusions 2 moves downward by the amount of low density particles 4 is moved upward.
遠心する場合の遠心力と時間は、分別しようとする微粒子の種類や量により適宜決めればよい遠心後は、固体介在物より上にある低比重微粒子4を含む液相を遠心管の傾斜により、又は、ピペットなどの吸引具により取り出すことにより、容易に高低二群の微粒子を分別することができる。 The centrifugal force and time when centrifuging, after centrifugation may be determined as appropriate depending on the type and amount of fine particles to be separated is a liquid phase containing a low specific gravity particles 4 in the above solid inclusions by the inclination of the centrifuge tube, or, by taking out by suction tool such as a pipette, it is possible to easily separate the fine particles of the high and low two groups.
【0009】 [0009]
【実施例】 【Example】
実施例1 Example 1
シリカ粉末で比重調節し加熱融解した寒天(比重1.05)3mlを予め内壁にパラフィンを塗布した15ml容の遠心管に注ぎ、冷却して固化させ、遠心管に内接する大きさの上面が遠心力ベクトル軸方向に対して約60度の斜面を形成している固体介在物を成形した。 Silica powder is poured into a centrifuge tube having a specific gravity adjusted by heating molten agar (specific gravity 1.05) 15 ml volume was applied pre inner wall paraffin to 3ml with, cooled and solidified, the size of the top surface inscribed in a centrifuge tube centrifuge the solid inclusions against a force vector axis direction to form a slope of about 60 degrees was formed. この遠心管にヒト末梢血(比重:約1.056)5mlを注ぎ、遠心装置にかけて300Gで、15分間遠心した。 The human peripheral blood in a centrifuge tube (specific gravity: about 1.056) Pour 5 ml, at 300G centrifuged apparatus and centrifuged for 15 min. 固体介在物の傾斜面の下部に沈降した赤血球(平均比重:1.096)が器壁を伝って固体介在物の下方に移動し、同時に固体介在物が上方に押し上げられた。 Solid inclusions of the inclined surface lower sedimentation erythrocytes (mean specific gravity: 1.096) of along the vessel wall to move downwards of the solid inclusions, solid inclusions is pushed upward at the same time. 最終的には固体介在物の上方に白血球と血漿成分(比重:1.0215)を含む画分が残り、下方に赤血球が集積した。 Eventually leukocytes and plasma component over the solid inclusions (specific gravity: 1.0215) remains fraction containing red blood cells integrated downward.
【0010】 [0010]
この実験の結果を〔表1〕に纏めた。 It summarizes the results of this experiment in Table 1.
実施例2 Example 2
固体介在物の遠心管内壁と接するポリスチレン樹脂製の球(比重1.05)を用い、500Gで20分遠心した外は実施例と同様に血液を処理した。 Using solid inclusions polystyrene resin spheres in contact with the centrifugal tube wall (specific gravity 1.05), the outer centrifugation 20 minutes at 500G was treated with Example as well as blood. その結果も〔表1〕に纏めた。 The result is also summarized in Table 1.
【0011】 [0011]
【表1】 [Table 1]
【0012】 [0012]
【発明の効果】 【Effect of the invention】
本発明の方法は、比較的簡単な装置を用いて、従来法の遠心分離操作を行うだけで、比重の異なる二種以上の微粒子を含む微粒子群を、比重の高低二群に高い精度で分画することができる。 The method of the present invention uses a relatively simple apparatus, only by performing the centrifugation of a conventional method, a group of fine particles containing two or more particles having different specific gravities, minutes with high accuracy to the level two groups of specific gravity it can demarcating. 特に、血液中から赤血球を分離し、有核細胞である白血球を摂取するのに極めて適した方法である。 In particular, red blood cells were separated from the blood, is very suitable way to ingest leukocytes are nucleated cells.
【図面の簡単な説明】 BRIEF DESCRIPTION OF THE DRAWINGS
【図1】傾斜した上面を有する固体介在物の平面図【図2】傾斜した上面を有する固体介在物の垂直断面図【図3】球状の固体介在物を配置した遠心管の垂直断面図【図4】遠心前の傾斜した上面を有する固体介在物を収容した遠心管垂直断面の模式図【図5】 遠心中の傾斜した上面を有する固体介在物を収容した遠心管垂直断面の模式図【図6】遠心終了時の傾斜した上面を有する固体介在物を収容した遠心管垂直断面の模式図【符号の説明】 [1] vertical sectional view of a centrifuge tube arranged vertically sectional view [FIG 3] Spherical solid inclusions solid inclusions having a plan view Figure 2 inclined upper surface of the solid inclusions having an inclined upper surface [ Figure 4 is a schematic view of a centrifuge tube vertical section containing the solid inclusions having a distal front of the inclined upper surface [5] a schematic view of a centrifuge tube vertical section containing the solid inclusions having an inclined upper surface in a centrifuge [ Figure 6 is a schematic view of a centrifuge tube vertical section containing the solid inclusions having an inclined upper surface at the end of the centrifugation [description of symbols]
1. 1. 遠心管2. Centrifuge tube 2. 固体介在物3. Solid inclusions 3. 赤血球4. Red blood cells 4. 有核細胞5. Nucleated cells 5. 固体介在物の傾斜した上面と遠心管内壁が漸近する部分6. Portion 6 sloping top and centrifuge tube wall of the solid inclusions is asymptotic. 遠心管底部7. Tube bottom 7. 遠心管胴部8. Centrifuge tube body portion 8. 固体介在物の微粒子群と接触する面9. Surface 9 in contact with the group of fine particles of solid inclusions. 低比重微粒子と高比重微粒子とが接する面10. Surface 10 of a low specific gravity particles and high density particles are in contact. 血液 blood

Claims (7)

  1. 比重の異なる二種以上の微粒子を含む微粒子群を、その微粒子群の比重の中間帯に位置する比重を持ち、微粒子群との接触面の外縁が遠心管に内接して上下に移動可能である固体介在物と共に遠心して、遠心管内壁と固体介在物との接触面を通して高比重微粒子を固体介在物の下方に、または低比重微粒子を固体介在物の上方に移動させると同時に固体介在物を比重の高低二粒子群の中間に移動、介在させることを特徴とする比重の異なる微粒子の分別法。 Microparticles group including two or more particles having different specific gravities, have a specific gravity which is located in the middle zone of the specific gravity of the microparticle group, the outer edge of the contact surface between the particulate group is movable vertically inscribed in a centrifuge tube by centrifugation with solid inclusions, centrifuge tube walls and solid inclusions specific gravity through the contact surface of high specific gravity particles below the solid inclusions, or low-density particulate solid inclusions simultaneously solid inclusions is moved upwardly in fractionation of fine particles having different specific gravity move to the middle of the high and low second grain group, and wherein the interposing of.
  2. 固体介在物の形状が、球状、楕円体状または微粒子群との接触面の少なくとも一部が遠心管内壁に対して鋭角で接触している柱状であることを特徴とする請求項1記載の方法。 The shape of the solid inclusions spherical method of claim 1, wherein at least a portion of the contact surface with the ellipsoidal or particulate groups characterized in that it is a columnar in contact at an acute angle to a centrifugal tube wall .
  3. 微粒子の粒径が0.01〜200μmの範囲内にあることを特徴とする請求項1記載の方法。 The method of claim 1, wherein the diameter of the particles is characterized in that in the range of 0.01~200Myuemu.
  4. 微粒子が液体中に懸濁しているかまたはコロイド状として存在していることを特徴とする請求項1記載の方法。 The method of claim 1, wherein the fine particles are present as either or colloidal suspended in a liquid.
  5. 微粒子を含む液体が、体液であることを特徴とする請求項1記載の方法。 Liquid containing fine particles The method of claim 1, wherein it is a body fluid.
  6. 固体介在物が、寒天、ゼラチン、合成樹脂、セラミクス、タンパク質、多糖類から選ばれた一種であることを特徴とする請求項1記載の方法。 Solid inclusions, agar, gelatin, synthetic resin, ceramic, protein, The method of claim 1, wherein it is a one selected from a polysaccharide.
  7. 遠心管内に、外縁が遠心管に内接し且つ上下に移動可能な固体介在物を収容したことを特徴とする請求項1記載の方法を実施するための装置。 A centrifuge tube, a device for carrying out the method of claim 1, wherein the outer edge containing a moveable solid inclusions inscribed and vertically into a centrifuge tube.
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